A5035332445- Protein Structure and Dynamics
- RNA Research and Splicing
- Virology and Viral Diseases
- RNA and protein synthesis mechanisms
- Enzyme Structure and Function
- Nuclear Structure and Function
- Click Chemistry and Applications
- Bacteriophages and microbial interactions
- Advanced Fluorescence Microscopy Techniques
- Respiratory viral infections research
- Advanced NMR Techniques and Applications
- Parvovirus B19 Infection Studies
- Chemical Synthesis and Analysis
- Viral Infections and Immunology Research
- Lipid Membrane Structure and Behavior
- Monoclonal and Polyclonal Antibodies Research
- Influenza Virus Research Studies
- Advanced Biosensing Techniques and Applications
- Biotin and Related Studies
- Advanced biosensing and bioanalysis techniques
- interferon and immune responses
- RNA modifications and cancer
- HIV Research and Treatment
- Mass Spectrometry Techniques and Applications
- Cellular transport and secretion
Leibniz-Forschungsinstitut für Molekulare Pharmakologie
2023-2025
Institut de Biologie Structurale
2015-2024
Centre National de la Recherche Scientifique
2015-2024
CEA Grenoble
2015-2024
Commissariat à l'Énergie Atomique et aux Énergies Alternatives
2015-2024
Université Grenoble Alpes
2015-2024
Université Paris-Saclay
2018
Université Paris-Sud
2018
European Molecular Biology Laboratory
2011-2017
European Molecular Biology Organization
2012-2015
Under tension: A set of genetically encoded unnatural amino acids can be used for biocompatible site-specific labeling proteins with fluorogenic dyes. The new compounds have norbornene and trans-cyclooctene units that react tetrazine derivatives in an inverse-electron-demand Diels–Alder cycloaddition (left picture). technique offers fast is orthogonal to through azide–cyclooctyne click reaction (right). Detailed facts importance specialist readers are published as ”Supporting Information”....
A bright click of life: Strained alkynes are genetically encoded into E. coli by use an engineered pyrrolysine amber suppressor tRNA/synthetase pair from Methanosarcina mazei. As a result cyclooctynyl lysine derivatives were incorporated site-specifically in proteins allowing efficient labeling with fluorogenic azide-bearing dyes means reaction under physiological conditions vitro and vivo (see scheme). The ability to visualize biomolecules within living specimen fluorescence tags has become...
The mechanisms by which intrinsically disordered proteins engage in rapid and highly selective binding is a subject of considerable interest represents central paradigm to nuclear pore complex (NPC) function, where transport receptors (NTRs) move through the NPC phenylalanine-glycine-rich nucleoporins (FG-Nups). Combining single-molecule fluorescence, molecular simulations, magnetic resonance, we show that rapidly fluctuating FG-Nup populates an ensemble conformations are prone bind NTRs...
Significance Conformational properties of unfolded and intrinsically disordered proteins (IDPs) under native conditions are important for understanding the details protein folding functions IDPs. The average dimensions these systems quantified using mean radius gyration end-to-end distance, measured by small-angle X-ray scattering (SAXS) single-molecule Förster resonance energy transfer (smFRET), respectively, although systematic discrepancies emerge from measurements. Through holistic sets...
Measles virus proteins form liquid droplets where they can encapsidate their genomic material.
Abstract Visualizing biomolecules by fluorescent tagging is a powerful method for studying their behaviour and function inside cells. We prepared genetically encoded an unnatural amino acid (UAA) that features bicyclononyne moiety. This UAA offered exceptional reactivity in strain‐promoted azide–alkyne cycloadditions. Kinetic measurements revealed the reacted also remarkably fast inverse‐electron‐demand Diels–Alder cycloaddition with tetrazine‐conjugated dyes. Genetic encoding of new...
Single-molecule methods have matured into central tools for studies in biology. Foerster resonance energy transfer (FRET) techniques, particular, been widely applied to study biomolecular structure and dynamics. The major bottleneck a facile general application of these arises from the need label biological samples site-specifically with suitable fluorescent dyes. In this work, we present an optimized strategy combining click chemistry genetic encoding unnatural amino acids (UAAs) overcome...
Nucleoporins (Nups), which are intrinsically disordered, form a selectivity filter inside the nuclear pore complex, taking central role in vital nucleocytoplasmic transport mechanism. These Nups display complex and nonrandom amino-acid architecture of phenylalanine glycine (FG)-repeat clusters intra-FG linkers. How such heterogeneous sequence composition relates to function could give rise mechanism is still unclear. Here we describe combined chemical biology single-molecule fluorescence...
Unter Spannung: Eine Reihe genetisch codierter nichtnatürlicher Aminosäuren kann für die biokompatible ortsspezifische Markierung von Proteinen mit fluorogenen Farbstoffen verwendet werden. Die neuen Verbindungen haben Norbornen- und trans-Cycloocten-Einheiten, Tetrazinen in einer Diels-Alder-Cycloaddition inversem Elektronenbedarf reagieren (links im Bild). Diese schnelle Markierungsreaktion ist orthogonal zur Azid-Cyclooctin-Klick-Reaktion (rechts).
Understanding the function of intrinsically disordered proteins is intimately related to our capacity correctly sample their conformational dynamics. So far, a gap between experimentally and computationally derived ensembles exists, as simulations show overcompacted conformers. Increasing evidence suggests that solvent plays crucial role in shaping has led several attempts modify water parameters thereby favor protein–water over protein–protein interactions. This study tackles problem from...
Intrinsically disordered proteins (IDPs) display a large number of interaction modes including folding-upon-binding, binding without major structural transitions, or through highly dynamic, so-called fuzzy, complexes. The vast majority experimental information about IDP have been inferred from crystal structures in complex with short peptides IDPs. However, provide mainly static view the complexes and do not give conformational dynamics experienced by bound state. Knowledge is fundamental...
Intrinsically disordered proteins (IDPs) are flexible biomolecules whose essential functions defined by their dynamic nature. Nuclear magnetic resonance (NMR) spectroscopy is ideally suited to the investigation of this behavior at atomic resolution. NMR relaxation increasingly used detect conformational dynamics in free and bound forms IDPs under conditions approaching physiological, although a general framework providing quantitative interpretation these exquisitely sensitive probes as...
Avian influenza polymerase undergoes host adaptation in order to efficiently replicate human cells. Adaptive mutants are localised on the C-terminal (627-NLS) domains of PB2 subunit. In particular, mutation residue 627 from E K rescues activity mammalian A transcription regulator ANP32A, comprising a long intrinsically disordered domain (IDD), is responsible for this adaptation. Human ANP32A IDD lacks 33 insertion compared avian and deletion restricts activity. We used NMR determine...
Studying the conformational landscape of intrinsically disordered and partially folded proteins is challenging only accessible to a few solution state techniques, such as nuclear magnetic resonance (NMR), small-angle scattering single-molecule Förster energy transfer (smFRET). While each techniques sensitive different properties chain, local structural propensities, overall dimension, or intermediate- long-range contacts, ensembles describing (IDPs) accurately should ideally respect all...
Gespannte Cycloalkine wurden in E. coli genetisch kodiert. Nach Modifikation des Pyrrolysyl-tRNA/Synthetase-Paars von Methanosarcina mazei konnten Cyclooctin tragende Lysinderivate positionsspezifisch durch Stop-Kodon-Unterdrückung Proteine eingebaut werden. Diese mit hohem Ertrag exprimiert und ermöglichten über eine Klick-Reaktion die effiziente Markierung fluorogenen Chromophoren (siehe Schema). Die Möglichkeit der Visualisierung Biomolekülen lebenden Objekten Fluoreszenzmarkierung ist...
Measles virus genome encapsidation is essential for viral replication and controlled by the intrinsically disordered phosphoprotein (P) maintaining nucleoprotein in a monomeric form (N) before nucleocapsid assembly. All paramyxoviruses harbor highly amino-terminal domains (PNTD) that are hundreds of amino acids length whose function remains unknown. Using nuclear magnetic resonance (NMR) spectroscopy, we describe structure dynamics 90-kDa N0PNTD complex, comprising 450 acids, at atomic...
Influenza A RNA polymerase complex is formed from three components, PA, PB1, and PB2. PB2 independently imported into the nucleus prior to reconstitution. All crystallographic structures of C-terminus (residues 536-759) reveal two globular domains, 627 NLS, that form a tightly packed heterodimer. The molecular basis affinity 627-NLS for importins remained unclear these structures, apparently requiring large-scale conformational changes importin binding. Using combination solution-state NMR,...
Deuterated oxazines are brighter and more photostable fluorophores in the near-infrared range amenable to cell imaging single molecule spectroscopy.
The acid stress chaperones HdeA and HdeB of Escherichia coli prevent the aggregation periplasmic proteins at acidic pH. We show in this report that they also form mixed aggregates with have failed to be solubilized pH allow their subsequent solubilization neutral HdeA, HdeB, together display an increasing efficiency for protein 3. They are less efficient 2, whereas is most efficient. Increasing amounts draw chaperone into pellets, suggesting co-aggregate substrate proteins. observed a...
Abstract Measles virus RNA genomes are packaged into helical nucleocapsids (NCs), comprising thousands of nucleo‐proteins (N) that bind the entire genome. N‐RNA provides template for replication and transcription by viral polymerase is a promising target inhibition. Elucidation mechanisms regulating this process has been severely hampered inability to controllably assemble NCs. Here, we demonstrate self‐organization N NC‐like particles in vitro upon addition RNA, providing simple versatile...